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American Journal of Pathology, Vol 138, 279-284, Copyright © 1991 by American Society for Investigative Pathology

REGULAR ARTICLES

Detection of loss of heterozygosity in formalin-fixed paraffin-embedded tumor specimens by the polymerase chain reaction

AB Bianchi, NM Navone and CJ Conti
Department of Carcinogenesis, University of Texas M.D. Anderson Cancer Center, Smithville 78957.

A polymerase chain reaction-based procedure was used for the detection of DNA length polymorphisms generated by naturally occurring genetic deletions or insertions of known sequence. This method consists of a simple one-step assay that does not require any restriction enzyme analysis or Southern blot hybridization, allowing identification in ethidium bromide-stained gels. The procedure described here was used to detect loss of heterozygosity at various loci, including the Hbb beta- globin gene cluster, in chemically induced mouse skin tumors, using a variety of tissue preparations, including microdissection of formalin- fixed, paraffin-embedded specimens, short-term cultures, and fluorescence-activated cell sorting of epithelial populations. This approach may be useful in detecting tumor-specific reduction to homozygosity at polymorphic chromosomal loci, allowing the mapping of putative tumor-suppressor loci involved in carcinogenesis.

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